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WO2008000846A1 - Procédé de stérilisation de substances à base de cacao à l'aide de co2 supercritique - Google Patents

Procédé de stérilisation de substances à base de cacao à l'aide de co2 supercritique Download PDF

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Publication number
WO2008000846A1
WO2008000846A1 PCT/ES2006/000360 ES2006000360W WO2008000846A1 WO 2008000846 A1 WO2008000846 A1 WO 2008000846A1 ES 2006000360 W ES2006000360 W ES 2006000360W WO 2008000846 A1 WO2008000846 A1 WO 2008000846A1
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WO
WIPO (PCT)
Prior art keywords
cocoa
sterilizing
cocoa material
contact
sterilized
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/ES2006/000360
Other languages
English (en)
Spanish (es)
Inventor
Elena Cienfuegos-Jovellanos Fernandez
Begoña MUGUERZA MARQUINEZ
Yolanda Castilla Escobar
Lourdes Calvo Garrido
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Laboratorio Reig Jofre SA
Original Assignee
Natraceutical SA
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Natraceutical SA filed Critical Natraceutical SA
Priority to PCT/ES2006/000360 priority Critical patent/WO2008000846A1/fr
Publication of WO2008000846A1 publication Critical patent/WO2008000846A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23G—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
    • A23G1/00—Cocoa; Cocoa products, e.g. chocolate; Substitutes therefor
    • A23G1/02—Preliminary treatment, e.g. fermentation of cocoa
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23B—PRESERVATION OF FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES; CHEMICAL RIPENING OF FRUIT OR VEGETABLES
    • A23B2/00—Preservation of foods or foodstuffs, in general
    • A23B2/70—Preservation of foods or foodstuffs, in general by treatment with chemicals
    • A23B2/704—Preservation of foods or foodstuffs, in general by treatment with chemicals in the form of gases, e.g. fumigation; Compositions or apparatus therefor
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23B—PRESERVATION OF FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES; CHEMICAL RIPENING OF FRUIT OR VEGETABLES
    • A23B2/00—Preservation of foods or foodstuffs, in general
    • A23B2/70—Preservation of foods or foodstuffs, in general by treatment with chemicals
    • A23B2/704—Preservation of foods or foodstuffs, in general by treatment with chemicals in the form of gases, e.g. fumigation; Compositions or apparatus therefor
    • A23B2/708—Preservation of foods or foodstuffs, in general by treatment with chemicals in the form of gases, e.g. fumigation; Compositions or apparatus therefor in a controlled atmosphere, e.g. partial vacuum, comprising only CO2, N2, O2 or H2O

Definitions

  • the present invention describes a method for the sterilization of cocoa-derived products that can be carried out using a supercritical fluid, particularly supercritical CO 2 .
  • BACKGROUND OF THE INVENTION According to the dictionary of the Royal Academy, the process of sterilizing is that which is carried out on an object to eliminate or destroy the possible pathogenic germs that it could have.
  • Sterilization is a treatment whose result is to preserve and keep the treated products unchanged for more or less long periods of time depending on the case.
  • the concept of sterility can be defined as the absence of all viable life forms, that is, the total absence of living microorganisms. A microorganism is alive from the microbiological point of view when it is capable of multiplying, therefore, a microorganism dies when it irreversibly loses the ability to reproduce.
  • the death of a microorganism is achieved when it is not possible to detect it in culture medium in which it has been shown to be able to proliferate. So the term sterility is expressed as the mathematical probability that a product remains contaminated with surviving microorganisms after being exposed to a sterilization process.
  • the European Pharmacopoeia sets a theoretical level of survival of no more than one microorganism in 10 6 sterilized units of final product.
  • the main thermal methods of food preservation are scalding, pasteurization and sterilization. Sterilization is a unit operation in which food is heated to a sufficiently high temperature (between 110 0 C and 14 0 0 C) and a sufficiently long time, to destroy the microbial activity to the maximum
  • Food Safety 9, 253 (1989) where they investigated the treatment in the elimination of natural microflora in some products such as flour, the components of pizza (cheese, salsa de tomato, fresh onions, bread,) or in ingredients such as rosemary, garlic powder and green pepper. They also investigated the method in whole fruits such as strawberries, melons and cucumber in order to delay the appearance of mold. The result was considered satisfactory although they observed partial destruction of fatty tissue.
  • GRAS GRAS solvent
  • thermo-labile nature due to the increasing demand that exists for functional foods, which in many cases incorporate one or more active principles of a thermo-labile nature, it is often necessary to use extraction and conservation techniques that operate at mild temperatures and avoid use of chemical agents that can leave residues unfit for human consumption. Moreover, if both processes can be carried out at the same stage, the process is greatly simplified and cheaper and meets the requirements of an environmentally sustainable process.
  • Supercritical CO 2 as a very suitable solvent to carry out extraction operations in cocoa products and surprisingly we have found that it is also the ideal solvent to carry out the sterilization of cocoa materials although in this case it requires the previous humidification of said materials of cocoa.
  • thermo-resistant microorganisms • It allows to eliminate spores of thermo-resistant microorganisms at much lower temperatures than those used in conventional sterilization methods (65 0 C vs 12O 0 C) and at much more moderate pressures than those used in traditional pasteurization methods (300 bar vs 3000 bar)
  • the sterilized cocoa product according to Ia contains 99.99% of the total sum of flavanols: monomers of catechin and epicatechin as well as dimers of procyanidins B1 and B2.
  • thermolabile active ingredients originally present or added (for example, vitamin C).
  • CO 2 has the following advantages over any of the other organic solvents: a) it does not need further disposal; b) leaves no residue in the product; c) it can isolate the compounds extracted selectively by changing the pressure, thus eliminating many separation and purification operations.
  • This invention describes a sterilization method by using CO 2 in a supercritical state for application to cocoa and its derivatives. More specifically, it refers to a method for sterilizing a cocoa material characterized in that it essentially comprises humidifying said cocoa material previously and then treating it with CO 2 in a supercritical state or in conditions close to the critical point, so that they are under pressure conditions and suitable temperature, and in contact for a sufficient period of time so that when CO 2 is separated, in said cocoa material, practically defatted in its entirety, a level of thermophilic aerobic thermo-resistant spores of less than 10 cfu / g is achieved, a mesophilic aerobic heat-resistant spore level less than 10 cfu / g and a mold and yeast level less than 10 cfu / g.
  • microorganisms present in the product that can be eliminated by the present invention are bacteria and their spores, fungi and their spores, yeasts, etc.
  • different enzymes could be deactivated, such as pectinase, amylase, proteinase, lipase, carboxypeptidase, etc.
  • sterilization is applied when a level of thermophilic aerobic thermo-resistant spores of less than 10 cfu / g is achieved, a level of mesophilic aerobic thermo-resistant spores less than 10 cfu / g and a level of molds and yeasts less than 10 cfu / g.
  • the viability of the cells is determined using techniques known in the art.
  • the starting product that forms the "cocoa material" of the present invention is Clone CCN-51 of the region of Quevedo (Ecuador), selected for its high polyphenol content.
  • Another important aspect with reference to the raw material for the elaboration of the cocoa material is that unfermented and unroasted seeds are used.
  • a process whereby a cocoa concentrate with a high polyphenol concentrate which comprises the steps of : a) subject the unfermented cocoa beans to a blanching in water at a temperature of 85-100 0 C for a time of 3 to 15 minutes; b) dry the seeds at a temperature not exceeding 85 0 C obtaining seeds with a humidity of no more than 15%; c) grind the seeds obtained in the previous stage until at least 99% of the product has a particle size of 300 ⁇ m; d) subject the ground seeds to extraction; and e) concentrate said cocoa extract with polyphenols; whose process can optionally include a degreasing stage that is performed before stage (d).
  • the extraction of cocoa can be carried out by means of supercritical CO 2 from cocoa with the original fat content or partially defatted by pressing, but in both cases to obtain the sterilization effect, the seed must be moistened previously with water.
  • the water content that the cake or cocoa powder must contain when being treated with supercritical CO 2 must be between 1 and 50% humidity, preferably between 5% and 40% humidity, so that the humidity of the cocoa product to be treated and further humidify it to the desired percentage of water for treatment with supercritical CO 2 .
  • the method to sterilize a cocoa material it can be made in various products, all obtained from variations in the different stages of the process of obtaining, but the preferred material is the powder obtained from the cake or wafer formed, once ground until a particle size of less than 500 microns, not having observed that the difference in size exerts influence on the sterilization of the product.
  • the process of sterilization of the cocoa material consists of putting it in contact with CO 2 under conditions of 50 to 500 bar pressure and a temperature of 20 0 C to 100 0 C. It has been determined that the flow of CO 2 must be between 5 / 2Og CO 2 / h / g of cocoa and 12Og CO 2 / h / g of cocoa, for a certain time that depends on the mentioned variables but can be approximately between 15 minutes and 6 hours.
  • a preferred embodiment of the invention is to start from a cake or cocoa powder that has been obtained from previously fermented seeds, peeled, pulped, blanched, dried and defatted by pressing, to sterilize it by contacting it with CO 2 a above 300 bar and a temperature of 65 0 C pressures, said cocoa material premoistened with 10% being water.
  • Another preferred embodiment is to contact said previously moistened cocoa material with 5% water and CO 2 at pressures greater than 300 bar and a temperature of 8O 0 C.
  • the sterilized cocoa obtained by the method of the invention has a thermophilic aerobic thermo-resistant spore level lower than 10 cfu / g, a mesophilic aerobic thermo-resistant spore level lower than 10 cfu / g and a mold and yeast level lower than 10 cfu / g.
  • cocoa powder sterilized by traditional means it has been inoculated with spores of Aspergillus niger and Aspergillus ochraceus, which have been removed by treating the cocoa powder with supercritical CO 2 according to the invention.
  • the cocoa obtained by the method of the invention once sterilized, can have a fat content equal to or less than 1%; the possible thermolabile compounds that were present at the beginning of the process, either naturally and / or added, they are not lost as a result of the process.
  • the content of total polyphenols (Folin-Ciocalteu measured) remaining in the sterilized powder is 99% with respect to the initial content, and in addition, the total sum of flavanols; Catechin and epicatechin monomers as well as procyanidin dimers B1 and B2 remaining in the sterilized powder is 99.99% with respect to the initial content.
  • cocoa powder as a preferred material are described below, although they could be applied to other cocoa products.
  • EXAMPLE 1 STERILIZATION OF COCOA POWDER
  • the product that has been used to exemplify the process has been an unfermented cocoa powder, which has been obtained from a previously peeled, pulped and scaled cocoa bean at 95 0 C for 5 minutes to inactivate the enzyme polyphenol oxidase, dried in a conventional dryer at a maximum temperature of 60 0 C, husked in a Lehmann equipment until a content of 2% by weight of residual husk on grain is achieved, and degreased by means of a mechanical pressing in continue using "expeliere" until you get a cocoa powder cake with a fat content of less than 12%.
  • the cocoa powder cake obtained has been stabilized at a temperature of 15 0 C, by passing it through a stainless steel vane cooler. This stabilized cocoa cake has been ground and sifted in a hammer mill until a ground cocoa powder is obtained with a particle size distribution of 99.99% lower than 500 microns.
  • thermophilic aerobes In the case of mesophilic aerobes, thermophilic aerobes, thermo-resistant mesophilic spores and thermo-resistant thermophilic spores, the sowing was performed on plates with DTPB agar (from English "Dextrose Tryptone Purple Bormocresol" - Scharlab), although the incubation temperature was 55 0 C and the incubation time of 48 hours for thermophilic microorganisms. The heat-resistant spore count was carried out after the successive dilutions were introduced in a bath of thermostated water at 8O 0 C. When the temperature in them reached that value, it was maintained for five minutes to eliminate viable bacteria and ensure that the count out only spores.
  • the efficacy of the treatment was determined from the residual cell viability data after the treatment that was quantified in cfu / g.
  • the ground raw material mother matter
  • Table 1 shows the results.
  • Table 1 Microbiological content of the original cocoa powder sample, without treatment. The results of the microbiological count and extraction efficiency performed in the cocoa powder samples treated with CO 2 after two hours of operation at different conditions are presented below. First, the equipment and the operation procedure are described.
  • the equipment used is shown in Figure 1. It consists of a CO 2 feed line, an extractor / sterilizer and a sampling system. CO 2 is fed liquid to vapor pressure (approx. 49 bar) after cooling in a thermostat bath at -10 0 C for not gasify during pumping is performed with a membrane pump (Milton Roy Dosapro) with refrigerated head A 100 cm long 316 stainless steel tube coil inserted in a heating blanket serves to preheat the CO 2 before entering the extractor / sterilizer, which is a pressure vessel (Robinson Mahoney micro-key reactor, Autoclave Engineers) of 316 stainless steel and 50 cm 3 capacity. The temperature control is carried out with a heating jacket connected to a thermocouple located on the external wall of the container.
  • a membrane pump Molton Roy Dosapro
  • a 100 cm long 316 stainless steel tube coil inserted in a heating blanket serves to preheat the CO 2 before entering the extractor / sterilizer, which is a pressure vessel (Robinson Mahoney micro-key
  • the temperature inside the sterilizer is measured with a thermocouple, while the pressure is monitored with a pressure gauge.
  • a safety valve set at 35 MPa and a rupture disk prevent the pressure from reaching values higher than those allowed.
  • the amount of CO 2 that passes per unit of time is determined in a mass flow meter connected to the end of the line.
  • the ground material is placed in the sterilizer forming a fixed porous bed in which redistributors or devices are inserted to favor contact and prevent the formation of preferred channels.
  • the heating jacket is connected and when the desired operating temperature has been reached, CO 2 begins to be pumped. Its temperature is adjusted as it passes through the preheater. Its inlet pressure and flow are regulated with the pump and with the valve. Once the pressure and temperature conditions have been adjusted, the CO 2 enters the bed at the bottom, and passes through it. The CO 2 together with the dissolved elements reaches the valve where the pressure is reduced until weather conditions.
  • the CO 2 goes to the gas state and the extracted materials precipitate (for example, the butter) and are collected in a previously tared tube. At fixed time intervals, the butter is weighed and the amount of CO 2 that has been necessary to use to extract said amount in the mass flow meter is read. Then it is released into the atmosphere.
  • FIG. 1 Equipment for sterilization-extraction with supercritical CO 2 of cocoa products.
  • Pl internal pressure.
  • Tl Indoor temperature.
  • the cocoa powder tended to be compacted inside the sterilizer, so that the CO 2 passed forming preferential channels fundamentally around the bed next to the wall of the extract leaving the outer part of a lighter color and the center darker showing that the butter It was only extracted in the periphery. This was manifested in a low efficiency of the reaction. Therefore, the use of various types of fillers and redistributors was explored. Its impact on the extraction curves is compared in Figure 2. As can be seen, the use of beads of approx. 3 mm of diameter with internal perforations and the use of redistributors made with stainless steel fine mesh were the most effective methods to allow the contact of CO 2 with the cake. In the industrial process, commercial fillers or redistributors could be used. But to operate in the laboratory, the simplest way was to introduce a redistributor every 2 cm of bed and thus proceeded for successive experiments.
  • the temperature In a non-thermal preservation method that is intended to be applied to thermolabile compounds, the temperature must be limited to a moderate range so as not to cause damage to the product to be treated. For that reason, the temperature that has been studied has oscillated between 40 and 8O 0 C. In this temperature range and varying the pressure between 150 and 300 bar it has not been possible to reduce the microbial content of the treated samples. On the other hand, it has been observed that the wetting of the raw material is necessary to achieve complete sterilization. Although the starting material already It contained a humidity of 8%, the effect that the increase in the initial moisture of the cocoa powder had on the treatment efficiency has been explored.
  • the cocoa powder was sprayed with different amounts of water (0, 10 and 25%), determining in each case the percentage of moisture obtained by weighing.
  • the 0% content corresponds to the starting material with the humidity it brought.
  • the samples so pretreated were subjected to the process with CO 2 at different temperatures and 300 bar. A 40 0 C regardless of the amount of water used (up to 25%) was not achieved reduce microbial counts.
  • the results obtained at higher temperatures are shown in Table 2.
  • the objective degree of sterilization is achieved, for example, by combining 8O 0 C and 5% of initial humidity, or at 65 0 C with 10% water. That is, only by combining a certain degree of humidity with a slightly elevated temperature, it is possible to reduce the microbiological content up to the conditions specified by the legislation for this type of products. It must be taken into account that the microbiological load of the raw material corresponds mainly to sporulated organisms and aerobic heat resistant. This explains why the temperature to be reached is at least 65 0 C, although with conventional methods, to eliminate this type of microorganisms it is necessary to exceed 12O 0 C.
  • the effect of the sterilization treatment with supercritical CO 2 on the content of total polyphenols as well as on the sum of its flavanols has been determined; the catechin and epicatechin monomers, and the procyanidin dimeros B1 and B2 Initially present in the treated cocoa powder.
  • the analyzes were performed on a sample of cocoa treated under the most extreme conditions and which could most influence the level of these phenolic compounds.
  • the conditions applied on the analyzed samples were 8O 0 C, 300 bar pressure with 10% water added.
  • EXAMPLE 2 DEACTIVATION OF SPORTS OF FUNGES Aspergillus niger And Aspergillus ochraceus POWDER POWDER

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  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • Food Preservation Except Freezing, Refrigeration, And Drying (AREA)

Abstract

L'invention concerne un procédé destiné à la stérilisation d'une substance à base de cacao à l'aide de CO2 supercritique, consistant principalement à humidifier au préalable ladite substance à base de cacao, puis à la traiter avec du CO2 à l'état supercritique ou dans des conditions proches du point critique, de façon qu'ils soient en contact pendant un laps de temps suffisant pour que, lors de la séparation du CO2, ladite substance à base de cacao, pratiquement dégraissée, ait une teneur en spores thermorésistantes aérobies thermophiles inférieure à 10 ufc/gr, en spores thermorésistantes aérobies mésophiles inférieure à 10ucf/gr et en champignons et en levures inférieure à 10 ucf/gr. Le traitement est réalisé à une pression comprise entre 50 et 500 bar, ce qui permet d'augmenter la vitesse d'extraction, à une température comprise entre 20 et 100°C et avec un débit compris entre 5 et 120 gr CO2/h/gr de cacao. La teneur restante en polyphénols totaux de la poudre stérilisée correspond à 99% de la teneur initiale.
PCT/ES2006/000360 2006-06-19 2006-06-19 Procédé de stérilisation de substances à base de cacao à l'aide de co2 supercritique Ceased WO2008000846A1 (fr)

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PCT/ES2006/000360 WO2008000846A1 (fr) 2006-06-19 2006-06-19 Procédé de stérilisation de substances à base de cacao à l'aide de co2 supercritique

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PCT/ES2006/000360 WO2008000846A1 (fr) 2006-06-19 2006-06-19 Procédé de stérilisation de substances à base de cacao à l'aide de co2 supercritique

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9466479B2 (en) 2005-02-28 2016-10-11 Oerlikon Metco Ag, Wohlen System and process for high-density, low-energy plasma enhanced vapor phase epitaxy

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005001059A2 (fr) * 2003-06-23 2005-01-06 Novasterilis Inc. Procede d'inactivation d'organismes en utilisant du bioxyde de carbone a pression et temperature critiques ou a leur voisinage
US20050084581A1 (en) * 2003-10-17 2005-04-21 Yoshiyuki Sato Apparatus for liquid food sterilization or enzyme deactivation with supercritical carbon dioxide, and method of liquid food sterilization or enzyme deactivation, and liquid food obtained by the use of the apparatus and the method
WO2005115160A1 (fr) * 2004-05-24 2005-12-08 Natraceutical, S.A. Procede permettant de produire un concentrat de polyphenol de cacao

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005001059A2 (fr) * 2003-06-23 2005-01-06 Novasterilis Inc. Procede d'inactivation d'organismes en utilisant du bioxyde de carbone a pression et temperature critiques ou a leur voisinage
US20050084581A1 (en) * 2003-10-17 2005-04-21 Yoshiyuki Sato Apparatus for liquid food sterilization or enzyme deactivation with supercritical carbon dioxide, and method of liquid food sterilization or enzyme deactivation, and liquid food obtained by the use of the apparatus and the method
WO2005115160A1 (fr) * 2004-05-24 2005-12-08 Natraceutical, S.A. Procede permettant de produire un concentrat de polyphenol de cacao

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
KAMIHIRA M. ET AL.: "Sterilization of microorganisms with supercritical carbon dioxide", AGRIC. BIOL. CHEM., vol. 51, no. 2, 1987, pages 407 - 412 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9466479B2 (en) 2005-02-28 2016-10-11 Oerlikon Metco Ag, Wohlen System and process for high-density, low-energy plasma enhanced vapor phase epitaxy

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